Journal: Journal of Biological Chemistry

Article Title: Plasminogen Activator Inhibitor-1 Regulates Tumor Growth and Angiogenesis

doi: 10.1074/jbc.m105980200

Figure Lengend Snippet: FIG. 3. Immuno-analysis of vessel architecture in M21 tumors. Tumor sections treated with PBS (A, D, G), partially inactivated PAI-1 (B, E, H), or fully active PAI-1 (C, F, I) were stained for the following: immunohistochemical staining for collagen IV (A–C), immunohisto- chemical staining for smooth muscle cell actin (D–F), immunofluores- cence analysis of vitronectin (green) (G–I), endothelial cells (red), and tumor cells (blue). The original magnification for all panels is 400.

Article Snippet: For immunofluorescence staining of tumor sections, slides were treated as above and then double stained with rabbit anti-murine vitronectin (Molecular Innovations) and rat anti-murine PECAM-1 (PharMingen) and detected with Flour 488 goat anti-rabbit IgG (Molecular Probes) and Texas Red rat anti-mouse IgG (Molecular Probes).

Techniques: Staining, Immunohistochemical staining

Journal: Veterinary Research

Article Title: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) moonlights as an adhesin in Mycoplasma hyorhinis adhesion to epithelial cells as well as a plasminogen receptor mediating extracellular matrix degradation

doi: 10.1186/s13567-021-00952-8

Figure Lengend Snippet: Binding of the rGAPDH protein to different ECM components in ELISA experiment . Microtiters plate was coated with Matrigel, fibronectin, collagen or laminin solution. Various concentrations of rGAPDH or BSA were added and detected by anti-His-tag monoclonal antibody. For detecting the binding to vitronectin, microtiters plate was coated with rGAPDH or BSA. Various concentrations of vitronectin were added and detected by anti-vitronectin monoclonal antibody. * P < 0.05, ** P < 0.01, compared with the negative control (BSA).

Article Snippet: The bound vitronectin was detected by rabbit anti-vitronectin monoclonal antibody (1:2000; Boster, China), followed by goat anti-rabbit IgG (1:10 000).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

doi: 10.3390/ijms12063409

Figure Lengend Snippet: Expression of integrin αv, β3 in different ovarian tissues.

Article Snippet: Rabbit anti-human integrin αv, β3 polyclonal antibody was purchased from Wuhan Boster Company, mouse anti-human Lewis y monoclonal antibody was purchased from Abcam Inc., biotinylated goat anti-mouse IgM, biotinylated goat anti-mouse IgG were purchased from Zhongshan Biotechnology Co., Ltd. Serum albumin (BSA) and DAB kit were purchased from Zhongshan Biotechnology Co., Ltd.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

doi: 10.3390/ijms12063409

Figure Lengend Snippet: Immunohistochemical staining in ovarian malignant tumor ( A1 – A3 ), borderline tumor ( B1 – B3 ), benign tumor ( C1 – C3 ) and normal ovarian tissue ( D1 – D3 ). Integrin αv ( A1 , B1 , C1 , D1 ); β3 ( A2 , B2 , C2 , D2 ) and Lewis y ( A3 , B3 , C3 , D3 ). (Original magnification ×200).

Article Snippet: Rabbit anti-human integrin αv, β3 polyclonal antibody was purchased from Wuhan Boster Company, mouse anti-human Lewis y monoclonal antibody was purchased from Abcam Inc., biotinylated goat anti-mouse IgM, biotinylated goat anti-mouse IgG were purchased from Zhongshan Biotechnology Co., Ltd. Serum albumin (BSA) and DAB kit were purchased from Zhongshan Biotechnology Co., Ltd.

Techniques: Immunohistochemical staining, Staining

Journal: International Journal of Molecular Sciences

Article Title: Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

doi: 10.3390/ijms12063409

Figure Lengend Snippet: Relationship between integrin αv with the clinical and pathological parameters of malignant ovarian cancer.

Article Snippet: Rabbit anti-human integrin αv, β3 polyclonal antibody was purchased from Wuhan Boster Company, mouse anti-human Lewis y monoclonal antibody was purchased from Abcam Inc., biotinylated goat anti-mouse IgM, biotinylated goat anti-mouse IgG were purchased from Zhongshan Biotechnology Co., Ltd. Serum albumin (BSA) and DAB kit were purchased from Zhongshan Biotechnology Co., Ltd.

Techniques: Diffusion-based Assay

Journal: International Journal of Molecular Sciences

Article Title: Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

doi: 10.3390/ijms12063409

Figure Lengend Snippet: Relationship between integrin β3 with the clinical and pathological parameters of malignant ovarian cancer.

Article Snippet: Rabbit anti-human integrin αv, β3 polyclonal antibody was purchased from Wuhan Boster Company, mouse anti-human Lewis y monoclonal antibody was purchased from Abcam Inc., biotinylated goat anti-mouse IgM, biotinylated goat anti-mouse IgG were purchased from Zhongshan Biotechnology Co., Ltd. Serum albumin (BSA) and DAB kit were purchased from Zhongshan Biotechnology Co., Ltd.

Techniques: Diffusion-based Assay

Journal: International Journal of Molecular Sciences

Article Title: Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

doi: 10.3390/ijms12063409

Figure Lengend Snippet: Correlation between expression of Lewis y antigen and integrin αv in ovarian cancer.

Article Snippet: Rabbit anti-human integrin αv, β3 polyclonal antibody was purchased from Wuhan Boster Company, mouse anti-human Lewis y monoclonal antibody was purchased from Abcam Inc., biotinylated goat anti-mouse IgM, biotinylated goat anti-mouse IgG were purchased from Zhongshan Biotechnology Co., Ltd. Serum albumin (BSA) and DAB kit were purchased from Zhongshan Biotechnology Co., Ltd.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

doi: 10.3390/ijms12063409

Figure Lengend Snippet: Correlation between expression of Lewis y antigen and integrin β3 in ovarian cancer.

Article Snippet: Rabbit anti-human integrin αv, β3 polyclonal antibody was purchased from Wuhan Boster Company, mouse anti-human Lewis y monoclonal antibody was purchased from Abcam Inc., biotinylated goat anti-mouse IgM, biotinylated goat anti-mouse IgG were purchased from Zhongshan Biotechnology Co., Ltd. Serum albumin (BSA) and DAB kit were purchased from Zhongshan Biotechnology Co., Ltd.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Study on the Expression and Clinical Significances of Lewis y Antigen and Integrin αv, β3 in Epithelial Ovarian Tumors

doi: 10.3390/ijms12063409

Figure Lengend Snippet: Integrin αv, β3 and Lewis y colocalize in ovarian malignant tumor. Using an immunofluorescent double-labeling method. Integrin αv and β3 ( A1 and B1 ); Lewis y ( A2 and B2 ); nucleus ( A3 and B3 ); Merged image ( A4 and B4 ). (Original magnification ×400).

Article Snippet: Rabbit anti-human integrin αv, β3 polyclonal antibody was purchased from Wuhan Boster Company, mouse anti-human Lewis y monoclonal antibody was purchased from Abcam Inc., biotinylated goat anti-mouse IgM, biotinylated goat anti-mouse IgG were purchased from Zhongshan Biotechnology Co., Ltd. Serum albumin (BSA) and DAB kit were purchased from Zhongshan Biotechnology Co., Ltd.

Techniques: Labeling

Journal: PLoS ONE

Article Title: Confocal Laser Scanning Microscopy Evaluation of an Acellular Dermis Tissue Transplant (Epiflex®)

doi: 10.1371/journal.pone.0045991

Figure Lengend Snippet: Summary of immunostaining with human antibodies against matrix components of the ADM; + means detectable by immunostaining, - means absence of any detectable signal.

Article Snippet: The following primary antibodies were used, each diluted 1∶100: rabbit anti-human collagen I (Rockland, USA), rabbit anti-human collagen II (Rockland, USA), rabbit anti-human collagen III (Abcam, UK), rabbit anti-human collagen IV (Rockland, USA), rabbit anti-human fibronectin (Abcam, UK), sheep anti-human hyaluronic acid (Biotrend, Germany), mouse anti-human laminin-5 (BD Bioscience, USA), rabbit anti-human laminin (Rockland, USA), mouse anti-human osteopontin (Santa Cruz Biotechnology, USA), mouse anti-human tenascin (NeoMarkers, USA), rabbit anti-human vitronectin (Biotrend, Germany), mouse anti-human thrombospondin-1 (Dianova, Germany).

Techniques: Immunostaining

Journal: The Journal of Biological Chemistry

Article Title: Repeating Structures of the Major Staphylococcal Autolysin Are Essential for the Interaction with Human Thrombospondin 1 and Vitronectin *

doi: 10.1074/jbc.M113.521229

Figure Lengend Snippet: Human TSP-1 binds preferentially to repeats R1ab-R2ab as shown by surface plasmon resonance studies. A, human TSP-1 (0.1 μg) was immobilized on 96-well plates (MaxiSorp) and incubated with various molecular ratios of AtlE R1ab-R2ab or AtlE R1ab. The binding of repeats was detected using a polyclonal anti-AtlE-R1ab-R2ab IgG followed by incubation with a peroxidase-coupled secondary antibody. Results are expressed as means ± S.D. (n = 3). **, p < 0.01; ***, p < 0.001; ns, not significant. B, surface plasmon resonance sensorgrams of heterologously expressed AtlE R1ab-R2ab show a dose-dependent binding to immobilized hTSP-1. A CM5 biosensor was coated with hTSP-1 (∼4000 response units), and the heterologously expressed repeats R1ab-R2ab of AtlE were used as analytes. The values of the control flow cells were subtracted from each sensorgram. C, surface plasmon resonance sensorgrams of heterologously expressed AtlE R1ab-2ab show a dose-dependent binding to immobilized human vitronectin. Vn was immobilized on the CM5 biosensor (∼2500 response units), and the heterologously expressed repeats R1ab-R2ab of AtlE were used as analytes. The values of the control flow cells were subtracted from each sensorgram. D, low binding activity of heterologously expressed AtlE repeat R1ab (25 μg/ml) to immobilized hTSP-1 as analyzed by an SPR study. Shown is an SPR sensorgram of a manual run.

Article Snippet: Contaminations with fibronectin or vitronectin were excluded by immunoblot analysis of purified hTSP-1with primary antibodies against vitronectin (rabbit anti-human vitronectin, CompTech) and fibronectin (rabbit anti-human fibronectin, Dako) and a secondary goat anti-rabbit IgG coupled to alkaline phosphatase (1:7500, Promega).

Techniques: SPR Assay, Incubation, Binding Assay, Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Repeating Structures of the Major Staphylococcal Autolysin Are Essential for the Interaction with Human Thrombospondin 1 and Vitronectin *

doi: 10.1074/jbc.M113.521229

Figure Lengend Snippet: Human TSP-1 and vitronectin bind dose-dependently to the R1ab-R2ab repeats of Atl and compete for binding. A, binding of hTSP-1 to immobilized Atl repeats R1ab-R2ab. The heterologously expressed repeats R1ab-R2ab (0.5 μg) were immobilized on microtiter plates (MaxiSorp) and incubated with increasing amounts of hTSP-1. B, binding of human vitronectin to immobilized Atl repeats R1ab-R2ab. The heterologously expressed repeats R1ab-R2ab (0.5 μg) were immobilized on microtiter plates (MaxiSorp) and incubated with increasing amounts of Vn. C and D, human TSP-1 competes with human vitronectin for binding to the immobilized Atl repeats R1ab-R2ab. The heterologously expressed repeats R1ab-R2ab (0.5 μg) were immobilized on microtiter plates (MaxiSorp) and incubated with hTSP-1 (1000 ng/well) in the presence of increasing molecular ratios of Vn (C) or with Vn (125 ng/well) in the presence of increasing molecular ratios of hTSP-1 (D). Bound hTSP was detected using a polyclonal mouse anti-hTSP-1 IgG antibody followed by incubation with a peroxidase-coupled secondary anti-mouse antibody, and bound Vn was detected using a polyclonal rabbit anti-Vn IgG followed by incubation with a peroxidase-coupled secondary anti-rabbit antibody. Results are expressed as means ± S.D. (n = 3). *, p < 0.05; ***, p < 0.001; ns, not significant.

Article Snippet: Contaminations with fibronectin or vitronectin were excluded by immunoblot analysis of purified hTSP-1with primary antibodies against vitronectin (rabbit anti-human vitronectin, CompTech) and fibronectin (rabbit anti-human fibronectin, Dako) and a secondary goat anti-rabbit IgG coupled to alkaline phosphatase (1:7500, Promega).

Techniques: Binding Assay, Incubation

Journal:

Article Title: Plasmin-Coated Borrelia burgdorferi Degrades Soluble and Insoluble Components of the Mammalian Extracellular Matrix

doi:

Figure Lengend Snippet: Degradation of soluble ECM components by plasmin-coated B. burgdorferi. The substrates tested were human fibronectin (A), laminin (B), vitronectin (C), collagen type I (D), collagen type III (E), and collagen type IV (F). Spirochetes were incubated in HBSS with no additions (B-UT) and with addition of uPA alone (B-uPA), PLG alone (B-PLG), and PLG and uPA together to form spirochete surface-associated plasmin (B-Plasmin). A sham preparation to control for free plasmin carryover in the latter group consisted of PLG and uPA in HBSS but no B. burgdorferi. ELISA plate wells (six replicates) coated with substrate were incubated for 6 h with 108 spirochetes from each experimental group. Undegraded substrate was detected, and percent degradation (a reduction in absorbance value was interpreted as substrate degradation) was calculated as described in Materials and Methods. Bars represent mean percent substrate degradation relative to the positive control (0% degradation) ± the standard deviation of six replicate wells for each experimental group. ∗, statistically significant (P < 0.0001); ∗∗, statistically significant (P < 0.01) compared to the positive control. The experiment was performed three times with similar results.

Article Snippet: Rabbit anti-human vitronectin and rabbit antibodies to human type I, III, and IV collagens were purchased from Biodesign International.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Positive Control, Standard Deviation

Journal:

Article Title: Plasmin-Coated Borrelia burgdorferi Degrades Soluble and Insoluble Components of the Mammalian Extracellular Matrix

doi:

Figure Lengend Snippet: Degradation of soluble ECM components by graded concentrations of plasmin-coated B. burgdorferi. The substrates tested were fibronectin (A), laminin (B), and vitronectin (C). Spirochetes were incubated in HBSS with no additions (B-UT) and with addition of PLG and uPA together to form spirochete surface-associated plasmin (B-Plasmin). A sham preparation to control for free plasmin carryover in the latter group consisted of PLG and uPA in HBSS but no B. burgdorferi. ELISA plate wells (six replicates) coated with substrate were incubated for 6 h with a range of plasmin-coated B. burgdorferi concentrations (106, 10 × 106, 50 × 106, and 100 × 106 per well) as well as 100 × 106 untreated spirochetes. Undegraded substrate was detected, and percent degradation (a reduction in absorbance value was interpreted as substrate degradation) was calculated as described in Materials and Methods. Bars represent mean substrate degradation relative to the positive control (0% degradation) ± the standard deviation of six replicate wells for each experimental group. ∗, statistically significant (P < 0.0001) compared to ELISA positive control. The experiment was performed twice with similar results.

Article Snippet: Rabbit anti-human vitronectin and rabbit antibodies to human type I, III, and IV collagens were purchased from Biodesign International.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Positive Control, Standard Deviation